Northern blot analysis of HvNAS1 gene expression in transgenic lines. The HvNAS1 ORF was used as a probe. Total RNA was extracted from the shoots of 44-day-old plants grown hydroponically under Fe-sufficient conditions, and 20 μg of total RNA was loaded into each lane. Ethidium bromide staining was used to ensure equal loading of rRNA. NT non-transgenic line; 35S, 35S-HvNAS1 transgenic line; AN1, AN2, and AN3, OsActin1-HvNAS1 transgenic lines.